Krüppel-like factor 10 (KLF10) regulates various cellular functions, such as proliferation, differentiation and apoptosis, as well as the homeostasis of several types of tissue. In the present study, we attempted a loss-of-function analysis of zebrafish Klf11a and Klf11b, which constitute human KLF10 homologs. Embryos injected with klf11b-morpholino (MO) showed developmental retardation and cell death, whereas klf11a-MO-injected embryos showed normal development. In klf11b-MO-injected embryos, a dramatic increase in the amount of zebrafish p53 mRNA might be the cause of the increase in that of bax. The degree of apoptosis decreased in the klf11b-MO and p53-MO co-injected embryos. These findings imply that KLF10 is a negative regulator of p53-dependent transcription, suggesting that the KLF10/p53 complex may play an important role in apoptosis for maintenance of tissue homeostasis during embryonic development.
Purpose: Of various effects of relaxin, we assumed that anti-fibrotic effects, neovascularization effects and vasodilatation effects of relaxin might enhance the survival rate of skin flap. In the current study, we used adenovirus expressing relaxin genes to examine whether these genes could enhance the survival rate of a skin flap. Methods: A total of 30 Sprangue-Dawley rats were divided into three groups: RLX group (10; relaxin virus injected group), CTR group (10; no gene coded virus injection group), and PBS group (10; PBS injected group). Each group was intradermally injected with the virus ($10^7$ PFU) and PBS 48 hours before and immediately before the flap elevation. A distally based flap $3{\times}9\;cm$ in size was elevated on the dorsal aspect of each rat. Following this, a flap was placed in the original location and then sutured using a #4-0 Nylon. A surviving area of the flap was measured and then compared on postoperative days 3, 7 and 10. Using a laser Doppler, the amount of blood flow was measured. On postoperative day 10, tissues were harvested for histologic examination and the number of blood vessels was counted. Results: There was a significant increase in the area of the flap survival in the RLX group on postoperative days 3 and 7. The Doppler measurement also showed significantly increased blood flow immediately after the operation and on postoperative days 7 and 10. The number of blood vessels was significantly greater in the RLX group in the tissue harvested on postoperative day 10. The VEGF concentration was significantly higher in the RLX group than others in the tissues harvested on postoperative day 10. Conclusion: Following an analysis of the effects of relaxin-secreting adenovirus on the survival of a flap, the surviving area of the flap and the blood flow also increased. A histopathology also showed an increase in the number of blood vessels and the concentration of VEGF.
Demineralized bone particle (DBP) has been used as one of the powerful inducers of bone and cartilage tissue specialization. In this study, we fabricated DBP/PLGA scaffold for tissue engineered disc regeneration. We manufactured dual-structured scaffold to compose inner cylinder and outer doughnut similar to nature disc tissue. The DBP/PLGA scaffold was characterized by porosity, wettability, and water uptake ability. We isolated and cultured nucleus pulposus (NP) and annulus fibrosus (AF) cells from rabbit intervertebral disc. We seeded NP cells into the inner core of the hybrid scaffold and AF cells into the outer portion of it. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl) -2,5- diphenyltetrazolium -bromide (MTT) test. PLGA and PLGA/DBP scaffolds were implanted in subcutaneous of athymic nude mouse to observe the formation of disc-like tissue in vivo. And then we observed change of morphology and hematoxylin and eosin (H&E). Formation of disc-like tissue was better DBP/PLGA hybrid scaffold than control. Specially, we confirmed that scaffold impregnated 20 and 40% DBP affected to proliferation of disc cell and formation of disc-like tissue.
Background Adipose tissue damage of cryopreserved fat after autologous fat transfer is inevitable in several processes of re-transplantation. This study aims to compare and analyze the survivability of adipocytes after thawing fat cryopreserved at $-20^{\circ}C$ by using thawing methods used in clinics. Methods The survival rates of adipocytes in the following thawing groups were measured: natural thawing at $25^{\circ}C$ for 15 minutes; natural thawing at $25^{\circ}C$ for 5 minutes, followed by rapid thawing at $37^{\circ}C$ in a water bath for 5 minutes; and rapid thawing at $37^{\circ}C$ for 10 minutes in a water bath. The survival rates of adipocytes were assessed by measuring the volume of the fat layer in the top layers separated after centrifugation, counting the number of live adipocytes after staining with trypan blue, and measuring the activity of mitochondria in the adipocytes. Results In the group with rapid thawing for 10 minutes in a water bath, it was observed that the cell count of live adipocytes and the activity of the adipocyte mitochondria were significantly higher than in the other two groups (P<0.05). The volume of the fat layer separated by centrifugation was also measured to be higher, which was, however, not statistically significant. Conclusions It was shown that the survival rate of adipocytes was higher when the frozen fat tissue was thawed rapidly at $37^{\circ}C$. It can thus be concluded that if fats thawed with this method are re-transplanted, the survival rate of cryopreserved fats in transplantation will be improved, and thus, the effect of autologous fat transfer will increase.
Purpose: Frostbite can affect still soldiers. Initial clinical manifestations are similar for superficial and deep frostbite, so early treatment is identical. It is under-estimated by physicians. We try to identify the challenges of managing these complex tissue injuries. Methods: A retrospective analysis of 84 patients hospitalized at AFCH from 2009 to 2015 was conducted. We investigated differences of epidemiological characteristics, identification of soft tissue injury, treatment and complications between superficial (SF: 43; 51.2%) and deep (DF: 41; 48.8%) frostbite. Results: The major (94.0%) developed frostbite in dry circumstances (89.3%). Wet circumstances (66.7%) were more susceptible to DF rather than dry (46.7%). The 38 (45.2%) arrived to specialist within 7days. Most prone sites were feet, followed by hands. Toes had more deep injuries. DF presented more increased levels of ALT, CPK, CKMB, CRP. The bone scan of W+S+ was 48.3%, 87.1% and W+S- was 20.7%, 12.9%, respectively. The treatment resulted in improved or normalized perfusion scan with matching clinical improvement. It was a good tool to assess treatment response. Eighteen normal and 8 stenotic type of PCR resulted in normal with matching clinical improvement. One continuous obstructive waveform led to minor amputation. Twelve underwent both PCR and MRA. Among 6 normal PCR, 5 showed normal and one stenosis in MRA. All 5 stenosis and one obstruction showed the same findings in MRA. It was a good tool to evaluate vascular compromise. They were treated with rapid rewarming (11.6%, 22.0%), hydrotherapy (16.3%, 29.3%), respectively. Six (14.6%) underwent STSG, 2 (4.9%) had digital amputation in DF. Berasil, Ibuprofen, Trental were commonly administered. PGE1 was administered selectively for 6.8, 10.8 days, respectively. Raynaud's syndrome (16.3%), CRPS (4.7%), LOM (14.6%) and toe deformity (4.9%) were specific sequelae. Conclusion: We should recommend intensive foot care education, early rewarming and evacuation to specialized units. The bone scanning and PCR should allow for a more aggressive and active approach to the management of tissue viability.
BACKGROUND/OBJECTIVES: The anti-diabetic activity of pear through inhibition of ${\alpha}-glucosidase$ has been demonstrated. However, little has been reported about the effect of pear on insulin signaling pathway in obesity. The aims of this study are to establish pear pomace 50% ethanol extract (PPE)-induced improvement of insulin sensitivity and characterize its action mechanism in 3T3-L1 cells and high-fat diet (HFD)-fed C57BL/6 mice. MATERIALS/METHODS: Lipid accumulation, monocyte chemoattractant protein-1 (MCP-1) secretion and glucose uptake were measure in 3T3-L1 cells. Mice were fed HFD (60% kcal from fat) and orally ingested PPE once daily for 8 weeks and body weight, homeostasis model assessment of insulin resistance (HOMA-IR), and serum lipids were measured. The expression of proteins involved in insulin signaling pathway was evaluated by western blot assay in 3T3-L1 cells and adipose tissue of mice. RESULTS: In 3T3-L1 cells, without affecting cell viability and lipid accumulation, PPE inhibited MCP-1 secretion, improved glucose uptake, and increased protein expression of phosphorylated insulin receptor substrate 1 [p-IRS-1, ($Tyr^{632})$)], p-Akt, and glucose transporter type 4 (GLUT4). Additionally, in HFD-fed mice, PPE reduced body weight, HOMA-IR, and serum lipids including triglyceride and LDL-cholesterol. Furthermore, in adipose tissue, PPE up-regulated GLUT4 expression and expression ratio of p-IRS-1 ($Tyr^{632})/IRS$, whereas, down-regulated p-IRS-1 ($Ser^{307})/IRS$. CONCLUSIONS: Our results collectively show that PPE improves glucose uptake in 3T3-L1 cells and insulin sensitivity in mice fed a HFD through stimulation of the insulin signaling pathway. Furthermore, PPE-induced improvement of insulin sensitivity was not accompanied with lipid accumulation.
The purpose of this study was to explore the expression of ATAD2 in ovarian tumor tissue as well as its relationship with degree of malignancy. Tumor tissue from 110 cases of ovarian cancer was collected in accordance with the Declaration of Helsinki for evaluation of ATAD2 expression iimmunohistochemistry, quantitative PCR (qPCR) and Western blotting. The correlation between the ATAD2 expression and and the prognosis of ovarian cancer was evaluated by Cox regression model. In addition, HO-8910 and OVCAR-3 cells were transfected with two siRNAs targeting ATAD2. Cell viability was evaluated with MTT assay, and cell migration by transwell migration assay. ATAD2 was shown to be highly expressed in 65.5% (72/110) of ovarian cancer cases, both at transcriptional and protein levels. Moreover, highly expression was positively correlated with degree of malignancy. Knock-down of ATAD2 in HO-8910 and OVCAR-3 cells was found to reduce cell migration. In addition, follow-up visits of the patients demonstrated that the 5-year survival rate was lower in patients with high expression of ATAD2. Our study suggested that ovarian tumor tissue may have highly expressed ATAD2, which is associated with tumor stage, omentum-metastasis, ascites and CA-125. Increased ATAD2 may play important roles in tumor proliferation and migration. ATAD2 could serve in particular as a prognostic marker and a therapeutic target for ovarian cancer.
Infections are caused due to the infiltration of tissue or organ space by infectious bacterial agents, among which Staphylococcus aureus bacteria are clinically most relevant. While current treatment modalities are in general quite effective, several bacterial strains exhibit high resistance to them, leading to complications and additional surgeries, thereby increasing the patient morbidity rates. Titanium dioxide is a celebrated photoactive material and has been utilized extensively in antibacterial functions, making it a leading infection mitigating agent. In view of the property amelioration in materials via nanofication, free-standing titania nanofibers (pure and nominally doped) and nanocoatings (on Ti and Ti6Al4V implants) were fabricated and evaluated to assess their efficacy to mitigate the viability and growth of S. aureus upon brief (30 s) activation by a portable hand-held infrared laser. In order to gauge the effect of exposure and its correlation with the antibacterial activities, both isolated (only titania substrate) and simultaneous (substrate submerged in the bacterial suspension) activations were performed. The bactericidal efficacy of the IR-activated $TiO_2$ nanocoatings was also tested against E. coli biofilms. Toxicity study was conducted to assess any potential harm to the tissue cells in the presence of photoactivated materials. These investigations showed that the photoactivated titania nanofibers caused greater than 97% bacterial necrosis of S. aureus. In the case of titania-coated Ti-implant surrogates, the bactericidal efficacy exceeded 90% in the case of pre-activation and was 100% in the case of simultaneous-activation. In addition to their high bactericidal efficacy against S. aureus, the benignity of titania nanofibers and nanocoatings towards tissue cells during in-vivo exposure was also demonstrated, making them safe for use in implant devices.
Purpose: The survival of bone marrow derived stem cell was reported several times. But the survival of adipose tissue derived stem cells(hASCs) was not mentioned on. We studied the adipose tissue derived stem cell's survival and effect on articular cartilage in rabbits. Methods: Osteoarthritis was induced in twenty New Zealand white rabbits by intraarticular injection of monosodium iodoacetate(MIA). After four weeks, hASCs were also injected into the knee joints space without any vehicle, but the control group received phosphate buffered saline only. The histologic grade of articular cartilage was measured in 4 and 8 weeks after the transplantation of hASC and the viability of injected stem cells measured by Fluorescent in situ Hybridization (FISH) examination. Results: After 4 and 8 weeks from hASCs transplantation, histologic grade was not significantly difference between two groups(p>0.05), and the Y chromosome of the transplanted hASCs was not detected in articular cartilage. Conclusion: We found that direct injection of hASC in joint space didn't work on damaged articular cartilage repair.
The obstacles for cardiac imaging are motion artifacts due to cardiac motion, respiration, and blood flow, and low signal due to small tissue volume of heart. To overcome these obstacles, fast imaging technique with ECG gating is utilized. Cardiac exam using MRI comprises of morphology, ventricular function, myocardial perfusion, metabolism, and coronary artery morphology. During cardiac morphology evaluation, double and triple inversion recovery techniques are used to depict myocardial fluidity and soft tissue structure such as fat tissue, respectively. By checking the first-pass enhancement of myocardium using contrast-enhanced fast gradient echo technique, myocardial blood flow can be evaluated. In addition, delayed imaging in 10 - 15 minutes can inform myocardial destruction such as chronic myocardial infarction. Ventricular function including regional and global wall motion can be checked by fast gradient echo cine imaging in quantitative way. MRI is acknowledged to be practical for integrated cardiac evaluation technique except coronary angiography. Especially delay imaging is the greatest merit of MRI in myocardial viability evaluation.
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